Chromatography

HPLC connections (TN #539)

This technical note shows possible types of connection between column and LC system. Bad connection may influence peak separation and should be avoided. The right connection is presented below.

How the Triple Quadrupole works?

The principle of Triple Quadrupole (TQ) is explained on EVOQ™ system by Bruker. The key points of the system are:

• Axial Ion Source
• Active Focusing Q1
• Lens-Free Ion Path
• 180° Collision Cell
• Elliptical Design
• Off Axis Detector

puriFlash

puriFlash^® Flash Cartridges

Interchim developed new technique for flash chromatography - Ultra Performance Flash Purification (UPFP) using special flash cartridges. The flash cartridges are in the form of regular or irregular silica. UPFP enables to run purifications with high purity of the yield and less solvent use.

Preparative LC

Task for preparative HPLC systems differs to analytical one. While analytical HPLC task is qualitative and quantitative determination of defined compounds in samples, preparative HPLC task is separation, purification and isolation of value products from mixtures.

Preparative chromatography can be devidet into three main areas:

• Semi-preparative separations
• Batch preparative chromatography (pilot or industrial scale)
• True Counter-current chromatography
• Simulated moving bed (SMB)
• Continuous chromatography

Scale definition

Preparative chromatography can be conbined with Flash chromatography in one system - purification device. The PuriFlash (Advion-Interchim) offers different modes of operation:

• Preparative line + Flash line
• Two Flash lines
• Two Preparative lines.

Introduction

Amino acids are key building blocks of life and play pivotal role in various metabolic pathways. They mostly act as intermediates often not directly involving proteins. Due to their chemical complexity and dynamic range, their reliable quantitative and qualitative analysis in biological fluids and tissues is crucial for nutritional information, compound identification and diagnostics.

For that purpose, a simple, elegant and thus far the most expeditious method for an invaluable amino acid analysis has been developed. LC/GC-MS based Metamino® kit offers comprehensive solution up to 75 metabolites including basic proteinogenic amino acids, biogenic amines and coenzymes with the possibility to a further analyte extension.

Features

• In principle suitable for any matrices (urine, blood serum, tear, cerebral liquid, tissue extracts, soil extracts, etc.)
• Easy sample preparation
• Sample derivatization and analysis under 20 minutes
• Broad portfolio of analytes (75) with the possibility for a further extension
• No sample heating/ freezing needed
• NIST Library for GC/MS available
• Possible to determine substances with a low molecular weight that can be degraded in the ion source (e.g., GLY, ALA, etc.)
• Suitable for the analysis of substances that are difficult to quantify, such as polyamines
• All necessary reagents, accessories, HPLC column and clear instructions for derivatization and analysis with high accuracy and sensitivity are included

Allowable Adjustments to HPLC Methods

Chromservis HPLC columns corresponding to USP and Eur. Pharmacopoeia methods and the extent to which the various parameters of a chromatographic test may be adjusted without fundamentally modifying the pharmacopoeial analytical procedures are listed in this technical note. Changes other than those indicated require revalidation of the procedure.

Sample loading in Flash chromatography

Liquid or solid, with small or large sample volumes: because your challenges are just as varied as your injections, we offer multiple possibilities for optimisation, to guarantee you the best results day after day.

Dry Load & Accessories

So far only available in a disposable plastic version, the Dry Loads are now also available in a stainless steel version. These are reusable and also have the advantage of greater resistance to pressure.
With the stainless steel Dry Loads, you can carry out solid injections on a preparative column and in flash column applications where the maximum pressure resistance of a plastic Dry Load would be insufficient.

Injection loops

Thanks to our range covering volumes from 100 μl to 50 mL, there is always an injection loop suited to your needs.

Autosampler

Automate your injections with our autosampler and increase your performance tenfold. The puriFlash® AS-1 allows the injection of samples from 500 µl to 500 mL, with an automated cleaning of the transfer tubes between each injection. The injection is managed by a sample queue on InterSoft software. The autosampler xan be equiped with 6-way or 10-way electric valve.

One rack/slot offers capacities fo the test tubes/bottles up to 250 mL and custom racks.

Injection Pump

When the quantity of product to be injected becomes important and when having to use a syringe and multiply the injections for a single purification becomes constraining, the injection pump is the ideal tool. You just have to prime the pump with the product and then launch the method. It’s as easy as that!

HPLC Phases

ASTRA - CHROMSERVIS

ARION - CHROMSERVIS

More information is available in Column care guide aswell.

CHROMSHELL - CHROMSERVIS

KINETEX - PHENOMENEX

* Columns are pH stable from 1.5 to 10 under isocratic conditions. Columns are pH stable from 1.5 to 8.5 under gradient conditions.

Kinetex 2.6µm columns with ID 2.1mm are pressure stable up to 1000 bar, otherwise up to 600 bar.

Kinetec chore-shell colums can be replace by new ChromShell colums. Just try it.

LUNA - PHENOMENEX

GEMINI - PHENOMENEX

SYNERGI - PHENOMENEX

ONYX - PHENOMENEX

JUPITER - PHENOMENEX

GraceSmart - GRACE

Alltech^® Prevail - GRACE

Nano / Capillary LC Column ProteCol - SGE

HOW TO CORRECTLY CRIMP VIALS

Crimp vials are excellent sample containers for automatic dispensers of gas and liquid chromatographs and for storing samples or calibration solutions. The technique of closing them is very important for proper tightness. Due to leakage caused by improper sealing, solvent evaporation or loss of analytes may occur.
A correctly closed vial can be recognized by the fact that its cap rotates with difficulty after closing and the septum is straight.
A vial that is closed with too much force can be recognized by the fact that its cap cannot usually be turned at all and, in addition, it has a bent septum (inwards). If the septum is punctured by the needle of the microsyringe, the septum will be heavily stressed and thus the vial's tightness will be compromised.
A vial that does not have a properly closed cap due to the low power of the crimping pliers is manifested by easy rotation of the cap and, in some cases, unfastened aluminum material around the lower edge of the vial neck.
You can set the correct force of the closing pliers.
In older types of pliers, the force is adjusted by turning the Allen key inside the jaws. Pliers also have a stop screw, which is used to set the safety distance, in order not to use too much force and thus to avoid leakage or even mechanical damage to the vial.